HSP22在肺缺血再灌注损伤中作用研究
发布时间：2019-03-18 14:04
【摘要】：[背景与目的]：在肺移植术、肺栓塞溶栓治疗等临床情况下,肺缺血再灌注损伤可引起急性肺功能不全,具有很高的发病率和死亡率。HSP22过表达在心肌缺血再灌注损伤中有保护作用,但其在肺缺血再灌注损伤中是否有保护作用未知。主要研究HSP22过表达在小鼠肺缺血再灌注损伤中的作用。 [方法]：采用WesternBlot方法检测HSP22转基因C57BL小鼠和野生型C57BL小鼠肺组织中HSP22的表达情况。取HSP22转基因C57BL小鼠12只,随机分为2组：假手术组、缺血再灌注组；野生型C57BL小鼠12只按相同方法分组：假手术组、缺血再灌注组。建立小鼠左肺原位缺血再灌注损伤模型。于缺血45分钟并再灌注120分钟后取动脉血行氧分压测定,留取左肺组织标本分别测定各组肺组织湿干重比值,丙二醛(MDA)含量,作光镜观察组织形态变化,免疫组织化学方法观察各组HSP22蛋白的表达情况,TUNEL法测定肺组织细胞凋亡的变化。 [结果]：WesternBlot法检测转基因小鼠肺组织中HSP22蛋白含量明显高于野生型小鼠组(P0.05),且表达量为野生型小鼠组的2.5倍。四组小鼠的动脉血氧分压无统计学差异(P0.05)。WT-I/R组小鼠的湿干重比值、肺泡损伤指数较WT-SO组显著升高(P0.01、P0.01), TG-I/R组小鼠的湿干重比值、肺泡损伤指数较TG-SO组明显升高(P0.05、P0.01),均提示转基因小鼠和野生型小鼠的肺缺血再灌注损伤模型成功建立。TG-I/R组小鼠的肺组织湿干重比值、MDA含量、肺泡损伤指数较WT-I/R组明显降低(均P0.05),提示过表达HSP22可减轻小鼠肺缺血再灌注损伤。免疫组化法示TG-I/R组小鼠HSP22积分光密度值较TG-SO组明显升高(P0.01),WT-I/R组小鼠HSP22积分光密度值较WT-SO组亦升高(P0.01),说明缺血再灌注损伤时HSP22表达上调。TUNEL法结果示：WT-I/R组较WT-SO组小鼠肺组织细胞荧光强度增强,TG-I/R组也较TG-SO组的荧光强度强,然而TG-I/R组较WT-I/R组荧光强度减弱,提示缺血再灌注损伤时细胞凋亡增加,但过表达HSP22可抑制损伤组织的细胞凋亡。 [结论]：1.HSP22转基因小鼠在F4代稳定传代,明确HSP22在转基因小鼠的肺组织内高表达。 2.在国内首次建立HSP22转基因小鼠肺缺血再灌注损伤模型。 3.过表达HSP22在肺缺血再灌注损伤中具有保护作用,可能的机制是HSP22抑制脂质过氧化和细胞凋亡。
[Abstract]:[background & objective]: in the clinical conditions of lung transplantation and thrombolytic therapy, pulmonary ischemia-reperfusion injury can cause acute pulmonary dysfunction. The overexpression of HSP22 has protective effect in myocardial ischemia-reperfusion injury, but it is unknown whether HSP22 has protective effect in lung ischemia-reperfusion injury. The aim of this study was to investigate the role of HSP22 overexpression in lung ischemia-reperfusion injury in mice. [methods]: WesternBlot method was used to detect the expression of HSP22 in lung tissue of HSP22 transgenic C57BL mice and wild type C57BL mice. Twelve HSP22 transgenic C57BL mice were randomly divided into two groups: sham-operation group and ischemia-reperfusion group, and wild-type C57BL mice were divided into two groups according to the same method: sham-operation group and ischemia-reperfusion group. The model of in situ ischemia reperfusion injury of left lung in mice was established. After 45 minutes of ischemia and 120 minutes of reperfusion, arterial blood was taken to measure the partial pressure of oxygen. The ratio of wet to dry weight of lung tissue and the content of malondialdehyde (MDA) in the lung tissue of each group were measured respectively, and the histological changes were observed under light microscope. Immunohistochemical method was used to observe the expression of HSP22 protein and TUNEL method was used to detect the changes of apoptosis in lung tissue. [results]: the content of HSP22 protein in lung tissue of transgenic mice was significantly higher than that of wild-type mice by WesternBlot (P0.05), and the expression level was 2.5 times higher than that of wild-type mice. There was no significant difference in arterial oxygen pressure among the four groups (P0.05) .Wet-dry weight ratio and alveolar injury index in WT-I / R group were significantly higher than those in WT-SO group (P 0.01, P0.01). The wet dry weight ratio and alveolar damage index in TG-I/R group were significantly higher than those in TG-SO group (P0.05, P0.01). The results showed that the model of lung ischemia-reperfusion injury in transgenic mice and wild-type mice was successfully established. The wet-dry weight ratio of lung tissue, the content of MDA and the index of alveolar injury in TG-I R group were significantly lower than those in WT-I/R group (P 0.05). It is suggested that over-expression of HSP22 can alleviate lung ischemia-reperfusion injury in mice. Immunohistochemical method showed that the integral optical density of HSP22 in TG-I/R group was significantly higher than that in TG-SO group (P0.01), and the integral optical density of HSP22 in WT-I/R group was also higher than that in WT-SO group (P0.01). The results of Tunel showed that the fluorescence intensity of lung cells in WT-I/R group was higher than that in WT-SO group, and the fluorescence intensity in TG-I/R group was higher than that in TG-SO group, and the fluorescence intensity of HSP22 in TG-I/R group was higher than that in TG-SO group. However, the fluorescence intensity of TG-I/R group was lower than that of WT-I/R group, suggesting that apoptosis increased during ischemia-reperfusion injury, but over-expression of HSP22 could inhibit the apoptosis of injured tissue. [conclusion]: 1.HSP22 transgenic mice were stably subcultured in F4 generation, and the high expression of HSP22 in lung tissue of transgenic mice was confirmed. 2. The model of lung ischemia-reperfusion injury in HSP22 transgenic mice was established for the first time in China. 3. Overexpression of HSP22 plays a protective role in lung ischemia-reperfusion injury. The possible mechanism is that HSP22 inhibits lipid peroxidation and cell apoptosis.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2442934
[Abstract]:[background & objective]: in the clinical conditions of lung transplantation and thrombolytic therapy, pulmonary ischemia-reperfusion injury can cause acute pulmonary dysfunction. The overexpression of HSP22 has protective effect in myocardial ischemia-reperfusion injury, but it is unknown whether HSP22 has protective effect in lung ischemia-reperfusion injury. The aim of this study was to investigate the role of HSP22 overexpression in lung ischemia-reperfusion injury in mice. [methods]: WesternBlot method was used to detect the expression of HSP22 in lung tissue of HSP22 transgenic C57BL mice and wild type C57BL mice. Twelve HSP22 transgenic C57BL mice were randomly divided into two groups: sham-operation group and ischemia-reperfusion group, and wild-type C57BL mice were divided into two groups according to the same method: sham-operation group and ischemia-reperfusion group. The model of in situ ischemia reperfusion injury of left lung in mice was established. After 45 minutes of ischemia and 120 minutes of reperfusion, arterial blood was taken to measure the partial pressure of oxygen. The ratio of wet to dry weight of lung tissue and the content of malondialdehyde (MDA) in the lung tissue of each group were measured respectively, and the histological changes were observed under light microscope. Immunohistochemical method was used to observe the expression of HSP22 protein and TUNEL method was used to detect the changes of apoptosis in lung tissue. [results]: the content of HSP22 protein in lung tissue of transgenic mice was significantly higher than that of wild-type mice by WesternBlot (P0.05), and the expression level was 2.5 times higher than that of wild-type mice. There was no significant difference in arterial oxygen pressure among the four groups (P0.05) .Wet-dry weight ratio and alveolar injury index in WT-I / R group were significantly higher than those in WT-SO group (P 0.01, P0.01). The wet dry weight ratio and alveolar damage index in TG-I/R group were significantly higher than those in TG-SO group (P0.05, P0.01). The results showed that the model of lung ischemia-reperfusion injury in transgenic mice and wild-type mice was successfully established. The wet-dry weight ratio of lung tissue, the content of MDA and the index of alveolar injury in TG-I R group were significantly lower than those in WT-I/R group (P 0.05). It is suggested that over-expression of HSP22 can alleviate lung ischemia-reperfusion injury in mice. Immunohistochemical method showed that the integral optical density of HSP22 in TG-I/R group was significantly higher than that in TG-SO group (P0.01), and the integral optical density of HSP22 in WT-I/R group was also higher than that in WT-SO group (P0.01). The results of Tunel showed that the fluorescence intensity of lung cells in WT-I/R group was higher than that in WT-SO group, and the fluorescence intensity in TG-I/R group was higher than that in TG-SO group, and the fluorescence intensity of HSP22 in TG-I/R group was higher than that in TG-SO group. However, the fluorescence intensity of TG-I/R group was lower than that of WT-I/R group, suggesting that apoptosis increased during ischemia-reperfusion injury, but over-expression of HSP22 could inhibit the apoptosis of injured tissue. [conclusion]: 1.HSP22 transgenic mice were stably subcultured in F4 generation, and the high expression of HSP22 in lung tissue of transgenic mice was confirmed. 2. The model of lung ischemia-reperfusion injury in HSP22 transgenic mice was established for the first time in China. 3. Overexpression of HSP22 plays a protective role in lung ischemia-reperfusion injury. The possible mechanism is that HSP22 inhibits lipid peroxidation and cell apoptosis.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
【参考文献】
相关博士学位论文 前1条
1 张付峰;HSP22转基因小鼠模型的建立[D];中南大学;2008年
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